南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (7): 1087-1092.doi: 10.12122/j.issn.1673-4254.2021.07.18

• • 上一篇    下一篇

近视小鼠巩膜成纤维细胞的基因表达谱：基于单细胞RNA测序的生物信息学分析

余嘉珍，莫 亚   

  • 出版日期:2021-07-20 发布日期:2021-07-19

Gene expression profiles of myopic mouse scleral fibroblasts: a bioinformatics analysis based on single-cell RNA sequencing

  • Online:2021-07-20 Published:2021-07-19

RichHTML

43

PDF

959

摘要： 目的 利用单细胞RNA测序( RNA-seq)技术探索巩膜成纤维细胞在近视形成前后基因表达的变化探索巩膜成纤维细胞在近视中的作用机制。方法 将正常健康的C57BL/6J小鼠按随机数字法平均分为近视模型组与阴性对照组，6只/组。阴性对照组双眼前节无遮盖、近视模型组使用半透明眼罩行右眼形觉剥夺，实验开始及结束时均测眼轴，并单细胞捕获右眼巩膜成纤维细胞进行RNA-seq，筛选差异表达基因，使用GO功能分析和京都基因和基因组百科全书（KEGG）途径进行功能显著性富集分析。结果 近视模型组与阴性对照组比较，筛选出差异基因169个（P>0.05），112个上调基因和 57个下调基因，分子功能中71个GO项（P<0.05）。GO功能分析显示差异基因中有：白细胞聚集、分化以及细胞黏附等与炎症相关GO项；ATP代谢与结合，氧化还原过程、氧化应激反应、氧化磷酸化等与缺氧相关的GO项；蛋白质折叠、蛋白质转运以及蛋白质代谢过程的负调控、内质网、内质网腔、内质网应激等与蛋白质及内质网应激相关GO的生物学过程。KEGG分析显示差异基因显著的富集信号通路主要为与缺氧相关的三羧酸循环、氧化磷酸化、过氧化物酶体增殖物激活受体信号通路、氧化磷酸化等通路，与炎症相关的丝裂原活化蛋白激酶信号通路、白细胞跨内皮转运等，以及与蛋白质相关的帕金森氏病、亨廷顿病、蛋白质消化吸收等通路。结论 巩膜成纤维细胞在近视形成过程中出现功能障碍，与炎症、缺氧、蛋白质调节及内质网应激的相关基因表达和通路的调节有一定关系。

关键词: C57BL/6J小鼠；形觉剥夺性近视；单细胞RNA-seq；巩膜成纤维细胞

Abstract: Objective To investigate the changes in gene expression profiles of mouse scleral fibroblasts after myopia using single-cell RNA sequencing technology and explore the mechanism of dysfunction of the scleral fibroblasts in myopia. Methods Normal healthy C57BL/6J mice were randomly divided into negative control group and myopia model group (n=6), and in the latter group, form deprivation myopia was induced in the right eye using translucent goggles. Single cell capture was performed in the right eye to obtain the scleral fibroblasts for RNA sequencing. The differentially expressed genes (DEGs) were screened and GO and KEGG analyses were carried out for functionally significant enrichment analysis. Results Comparison of the gene expression profiles identified a total of 169 DEGs between the myopia model group and the negative control group (P>0.05), including 112 up-regulated and 57 down-regulated genes. GO function analysis showed that the DEGs were involved in leukocyte aggregation, differentiation and adhesion and other inflammation-related terms; ATP metabolism and binding, redox process, oxidative stress response, oxidative phosphorylation and other GO terms related to hypoxia; protein folding, protein transport, negative regulation of protein metabolism, endoplasmic reticulum (ER), ER cavity, ER stress and other biological processes related to protein and ER stress. KEGG analysis analysis showed that the significantly enriched pathways of the DEGs involved mainly the TCA cycle, oxidative phosphorylation, PPAR signaling, oxidative phosphorylation and other pathways related to hypoxia; MAPK signaling pathway related to inflammation; leukocyte transendothelial transport; and protein-related Parkinson's disease, Huntington's disease, protein digestion and absorption pathways. Conclusion The dysfunction of the scleral fibroblasts occurs in myopia through complex mechanisms involving inflammation, hypoxia, protein regulation, and ER stress-related gene expression and pathway regulation.

Key words: C57BL/6J mice; form deprivation myopia; single-cell RNA sequencing; scleral fibroblasts